A New Species Of Bothriocephalus (Cestoda: Bothriocephalidae) From Lepomis Spp. (Actinopterygii: Centrarchidae) In North America Author Choudhury, Anindo Author Scholz, Tomáš Author Beuchel, Joseph S. text Journal of Parasitology 2022 2022-08-04 108 4 343 352 http://dx.doi.org/10.1645/21-70 journal article 10.1645/21-70 1937-2345 7746757 Bothriocephalus kupermani n. sp. ( Figs. 1–4 ) Informal synonyms: ‘ Bothriocephalus sp. ’ (PBI-487) of Brabec et al. (2015) ; ‘Pumpkinseed morph’ of Choudhury and Scholz (2020). Material studied: The description is based on whole, unstained, and unmounted specimens, specimens processed for SEM, and 18 specimens prepared as stained whole mounts. Measurements are from stained whole mounts as follows (number of worms in parentheses): 8 gravid specimens and 1 mature non-gravid specimen from pumpkinseed from Paul Lake, 5 November 2001 (2), Peter Lake, 13 July 2003 (4). Bolger Bog, 12 August 2003 (2) and Caribou Lake (1), and 1 gravid specimen from green sunfish from Duck Creek on 12 July 2009 . In addition, 8 stained immature worms from pumpkinseed from Paul Lake, Peter Lake, and Shawano Lake were also studied but not used for measurements. Description: Bothriocephalidea , Bothriocephalidae . Body 21– 43 mm ( 35 mm ) long, maximum width 973–1,595 (1,078) in posterior third. Strobila gradually widening posteriorly, comprising craspedote proglottids (primary and secondary), trapezoidal especially in anterior half, posterior proglottids with rounded edges, often losing craspedote appearance ( Figs. 1 , 2 ); proglottids covered with coniform spinitriches ( Fig. 4F, G ). Except for first few proglottids, immature proglottids including those without obvious anlagen of gonads numerous, wider than long, mature proglottids (with sperm in external sperm duct) very few (1–2), 175–507 (495) long by 781–1,232 (894) wide, length:width ratio 0.11–0.55 (0.55). Gravid proglottids much wider than long ( Fig. 1D, E ), first gravid proglottid 183–512 long by 750–1530 wide, length:width ratio 0.15–0.55, proglottids often narrower at end of strobila, posterior-most proglottid 252–750 (750) long by 751– 1,208 (751) wide, length:width 0.2–1.0 (1.0). Longitudinal musculature well-developed, formed by numerous muscle fibers. Osmoregulatory canals usually in several (usually 2–3) pairs ( Fig. 1E ). Ventral osmoregulatory canals wide, sinuous to almost straight ( Fig. 1E ). Dorsal osmoregulatory canals narrow, thin-walled, slightly sinuous, median to ventral osmoregulatory canals ( Fig. 1E ). Scolex 414–805 (458) long, with nearly parallel margins through most of its length, dorsoventrally ( Figs. 1B, C , 2 A, D , 3A, B ) and laterally ( Figs. 1A , 2C , 3C , 4B ), occasionally slightly wider in middle ( Figs. 1A , 3C , 4A ), maximum lateral width 151–210, maximum dorsoventral width 183–296 (296). Scolex gently narrowing anteriorly before widening to form slightly convex (almost blunt in dorsoventral view), bilobed apical disc with rounded outer corners ( Figs. 1A–C , 3A–C , 4A– C ). Apical disc 151–215 wide laterally, 179–234 (234) wide dorsoventrally. Bothria shallow, wide, opening anteriorly to shallow hollow at base of apical disc ( Figs. 1B, C , 3A, B , 4A ). Lateral surface of scolex may possess medial, shallow groove ( Figs. 4 A, B ). Neck short, 105–180 (180) wide, almost as wide as scolex and first proglottid ( Figs. 1A–C , 2A–D ). Scolex with coniform spinitriches, capilliform filitriches, tumuli ( Fig. 4A, D ), patch/tuft of capilliform filitriches at center of apical disc ( Fig. 1C, E ). Testes oval to subspherical, variable in size, 36–83 long by 26– 57 wide, medullary, 38–62 (49) per proglottid, ovoid to spherical, in 2 lateral fields on each side; most testes in main external (outer) field between ventral osmoregulatory canals, with only few testes in inner, narrow irregular field, median to internal ventral osmoregulatory canal ( Figs. 1E , 3F ). Cirrus sac median, anterior to ovary, transversely oval to subspherical in dorsoventral view, 61–74 (74) long by 63–90 (78) wide, leading to common genital pore on mid-dorsal surface of proglottid ( Fig. 1E ); genital pore postequatorial to almost equatorial, situated at 51–65% of proglottid length; external sperm duct (vas deferens) lateral to cirrus sac ( Fig. 1E ). Ovary compact, median, near posterior margin of proglottid, irregular in shape (slightly asymmetrical), weakly bilobed, with narrower mid-region (ovarian isthmus), 62–100 (88) long by 218– 368 (248) wide ( Figs. 1E , 3E ), occupying 23–38% of proglottid width. Vagina tubular, narrower in proximal part, sinuous, widened, thick-walled in distal part, opening to common genital pore on dorsal surface ( Fig. 1E ). Vitelline follicles very variable in shape, from elongate to almost spherical, 25–61 long by 20–46 wide, cortical, extensive, forming wide lateral fields on each side of proglottid from its lateral margins to beyond dorsal osmoregulatory canals medially, also variably and more sparsely present medially, often confluent by narrow field at level of ovary and uterine pore ( Figs. 1D, E , 3D ). Uterus tubular, thick-walled, looped in proximal part (anterodorsal to ovary and dorsal to vagina), then turning anterolaterally, circumventing vaginal canal ventrally, thereafter directed anteriorly, winding ( Fig. 1E ). In gravid proglottids, terminal (distal) part enlarged to form transversely oval, thick-walled uterine sac filled with numerous eggs; width of uterine sac in last proglottids 44–69% of proglottid width (usually about 50%). Uterine sacs open on ventral surface by almost medially situated, slit-like uterine pore situated at variable distance (at 18–37% of proglottid length) from anterior margin of proglottids ( Figs. 1D, E ). Eggs ovoid, 46–54 long by 32–34 wide (n ¼ 48), tanned when fully developed, operculate, unembryonated in utero. Table I. Museum specimens of Bothriocephalus from Lepomis spp. examined in this study and their redetermination.

Museum	Published
Original	accession	Date of	Determination	reference
designation	number	Host	Location	collection	Collector/author	(this study)	(where available)
Bothriocephalus	USNM 1391705	Lepomis macrochirus	Wisconsin	21 Jun 1944	D. Dunnel and D. Swanson. In	Bothriocephalus
claviceps	R.V. Bangham collection	kupermani
Bothriocephalus	USNM 1391706	L. macrochirus	‘‘Farm pond’’ Alabama	9 Apr 1955	R. V. Bangham collection	B. kupermani
claviceps
Bothriocephalus	USNM 1391707	Lepomis gibbosus	Tepee Lake, Wisconsin	Jun 1945	‘‘J.N.V.’’ in R.V. Bangham	B. kupermani
claviceps	collection
Bothriocephalus	USNM 1391708	L. gibbosus	Tepee Lake, Wisconsin	Jul 1945	‘‘M.M.’’ in R.V. Bangham	B. kupermani
claviceps	collection
Bothriocephalus	USNM 1391709	L. gibbosus	Merchants Lake, Algonquin Park,	21 Jul 1942	R. V. Bangham	B. kupermani	Bangham and
claviceps	Ontario, Canada	Venard (1946)
Bothriocephalus	USNM 1391712	L. gibbosus	White Trout Lake, Algonquin	6 Aug 1942	R. V. Bangham	B. kupermani	Bangham and
claviceps	Park, Ontario, Canada	Venard (1946)
Bothriocephalus	USNM 1391713	L. gibbosus	White Trout Lake, Algonquin	1942	R. V. Bangham	B. kupermani	Bangham and
claviceps	Park, Ontario, Canada	Venard (1946)
Bothriocephalus	USNM 1391714	L. gibbosus	Nebish Lake, Wisconsin	17 Jul 1948	R.V. Bangham	B. kupermani	Bangham (1944)
claviceps
Bothriocephalus	USNM 1348363	L. gibbosus	Walnut Lake, Michigan	13 May 1906	‘‘E.L.M.’’ Identified by A.R.	B. kupermani
claviceps	Cooper
Bothriocephalus	USNM 1358986	Lepomis cyanellus	Ottawa Co., Ohio	16 Jun 1970	M.C. Mondl/M.C. Mondl and	B. kupermani
claviceps	F.C. Rabalias
Bothriocephalus sp.	HWML 19744	L. cyanellus	Beckius Pond, Keith Co.,	1988	L. Larson	B. kupermani
Nebraska
Bothriocephalus sp.	HWML 19900	L. cyanellus	Beckius Pond, Keith Co.,	1988	M. Parsons	B. kupermani
Nebraska
Bothriocephalus	HWML 19929	L. cyanellus	Christensen Pond, Keith Co.,	28 Jul 1981	S.V.M.	B. kupermani
claviceps	Nebraska
Bothriocephalus	HWML 20414	L. macrochirus	Farm Pond -2, SE Lincoln,	25 May 1972	Monty Mayes	B. kupermani
claviceps	Lancaster Co., Nebraska
Bothriocephalus	HWML 30499	L. cyanellus	Buncombe Creek at Buncombe	14 May 1960	J. McDaniel, J. Teague Self	B. kupermani
claviceps	bridge, Hwy 32 Oklahoma	collection
Bothriocephalus	HWML 30500	Lepomis megalotis	Lake Texoma, University of	30 March 1959	J. McDaniel, J. Teague Self	B. kupermani
claviceps	Oklahoma Biological Station	collection
Boathouse, Marsall Co.,
Oklahoma
Bothriocephalus	HWML 30501	L. cyanellus	Buncombe Creek at Buncombe	14 May 1960	J. McDaniel, J. Teague Self	B. kupermani
claviceps	bridge, Hwy 32 Oklahoma	collection

Figure 1. Bothriocephalus kupermani n. sp. from Lepomis gibbosus . ( A ) Scolex and anteriormost proglottids in lateral view (Peter Lake). ( B, C ) Scoleces and anteriormost proglottids in dorsoventral view (B from Paul Lake, C from Shawano Lake). ( D ) Distribution of ventral vitelline follicles in two last mature and first gravid proglottids (note presence of median follicles connecting both lateral fields of vitelline follicles); holotype. ( E ) Gravid proglottid, dorsal view; only most lateral and most median vitelline follicles illustrated on dextral side of proglottid. Figure 2. Bothriocephalus kupermani n. sp. from Lepomis gibbosus . Anterior region of worms from different geographical locations showing features of the scolex and strobila. ( A ) Paul Lake (dorsoventral view), ( B ) Peter Lake (dorsolateral view), ( C ) Peter Lake (lateral view), and ( D ) Shawano Lake (dorsoventral view). Color version available online. Taxonomic Summary Type host: Lepomis gibbosus (Linnaeus, 1758) (Centrarchiformes: Centrarchidae ). Additional hosts: Lepomis cyanellus Rafinesque, 1819 (this study and museum material), L. macrochirus Rafinesque, 1819 (museum material), L. megalotis (Rafinesque, 1820) (museum material) (all Centrarchiformes: Centrarchidae ) (see Table I ). Site of infection: Intestine. Type locality: Paul Lake ( 46°15 ′ 04.78 ′′ N , 89°30 ′ 12.94 ′′ W ; 516 meters above sea level [m a.s.l.]), University of Notre Dame Environmental Research Center ( UNDERC ), Michigan ( Upper Peninsula ) near Land-o-Lakes, Wisconsin . Additional localities: Peter Lake ( 46°15 ′ 10.17 ′′ N , 89°30 ′ 12.94 ′′ W ; 518 m a.s.l.); Bolger Bog ( 46°13 ′ 47.37 ′′ N, 89°29 ′ 96 ′′ W ; 507 m a.s.l.), both UNDERC lakes, Wisconsin ; Shawano Lake , Wisconsin ( 44°47 ′ 11.88 ′′ N , 88°32 ′ 39.60 ′′ W ) ; Caribou Lake , Minnesota ( 47°42 ′ 59.04 ′′ N , 90°39 ′ 18.18 ′′ W ) , Duck Creek near Hobart , Wisconsin ( 44°32 ′ 01.09 ′′ N , 88°07 ′ 44.89 ′′ W ) , Fox River at Voyageur Park , De Pere , Wisconsin ( 44°27 ′ 03.39 ′′ N , 088°03 ′ 51.83 ′′ W ) . Geographical distribution: Canada ( Ontario ) , Mexico (precise locality not known; fish host introduced), United States ( Alabama, Michigan, Minnesota, Nebraska, Ohio, Oklahoma, Wisconsin ) . Infection rate: Mean abundance: 20.6 ± 25.3; 0–112 tapeworms/fish; prevalence: 83% of 30 fish examined from the type locality. Type material (all from L. gibbosus ): Holotype : USNM 1593369 , Paul Lake , UNDERC , Michigan , 5 November 2001 . Paratypes : USNM 1593370–1593373 , HWML 216811–216817 , IPCAS C-914 /1. Molecular data: The following DNA sequence data are available in GenBank (https://www.ncbi.nlm.nih.gov/genbank/) (the region of the genome is followed by the GenBank accession number: 18S rDNA ( ssr DNA): KR780953, 18S rDNA ( V 8 region, partial): MT 532462– MT 532463; ITS-1 (partial): MT 532532– MT 532537; ITS-2 (partial): MT 532512– MT 532513; 28S rDNA ( lsr DNA): KR780906, 16S rDNA ( rrnL ): KR780867; cytochrome c oxidase subunit 1 ( COI ): KR780814. ZooBank registration: urn:lsid:zoobank.org:act: 701C450F-6CE8-4CF8-ADB6-164F49BBAF39 . Etymology: The species is named after the late Boris Kuperman, whose sudden demise in 2002 robbed parasitology of a wise, gifted, and gentle practitioner who continued his new life in the United States with the same lifelong passion he always had for our beloved subject, and whose seminal work on the genus Triaenophorus inspired the senior authors (A.C. and T .S.) many years ago. Remarks Bothriocephalus kupermani differs from other species of Bothriocephalus parasitizing freshwater fishes in North America, namely B. claviceps , B. cuspidatus , B. formosus , and B. pearsei , by the following characteristics: (1) the scolex is almost rectangular in dorsoventral and lateral profiles, with nearly parallel margins (vs. arrow-shaped and narrowing towards the apical disc in B. cuspidatus and with maximum width in the anterior third in B. claviceps ), with shallow and wide bothria with almost parallel margins (widest in the posterior or middle part in other taxa); (2) small body size (maximum of 43 mm vs. 133 and 137 mm in B. cuspidatus and B. claviceps , respectively; Scholz, 1997 ); and (3) extensive vitelline follicles that are also present medially, i.e., in the cortex of the ovarian and uterine region (absent anteriorly and never or seldom confluent posteriorly in other species except B. formosus , which has markedly different scolex and tiny strobila compared to the new species; see Scholz, 1997 ; Choudhury and Scholz, 2020). Bothriocephalus kupermani is also typified by a relatively high number of immature proglottids without obvious anlagen of gonads and fewer testes (38–62) than B. cuspidatus from Walleye (62–101). Figure 3. Bothriocephalus kupermani n. sp. from Lepomis gibbosus . Scoleces and proglottids. ( A ) From Paul Lake (dorsoventral view), ( B ) from Shawano Lake (dorsoventral view), ( C ) from Peter Lake (lateral view). ( D ) Vitelline follicles, from Caribou Lake. ( E ) Ovary, from Caribou Lake. ( F ) Testes, from Peter Lake. Color version available online. The validity of B. kupermani is also supported by molecular data of Brabec et al. (2015 ; isolate called ‘ Bothriocephalus sp. n. , ‘PBI-487 ′ in their fig. 2) and Choudhury and Scholz (2020). The specimen from green sunfish, L. cyanellus , is similar in its morphology to specimens from the type host, L. gibbosus , and is considered conspecific.